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자료유형
학술저널
저자정보
저널정보
대한의생명과학회 대한의생명과학회지 대한의생명과학회지 제11권 제3호
발행연도
2005.9
수록면
327 - 332 (6page)

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Sf21 cells become popular as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains 0.1% D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike human glucose transporters, very little is known about the characteristics of the endogenous sugar transporter(s) in Sf21 cells. Thus, some kinetic properties of the sugar transport system were investigated, involving the uptake of 2-deoxy-D-glucose (2dGlc). In order to obtain a true measure of the initial rate of uptake, the uptake of [<sup>3</sup>H]2dGlc from both low (100μM) and high (10 mM) extracellular concentrations was measured over periods ranging from 30 sec to 30 min. The data obtained indicated that the uptake was linear for at least 2 min at both concentrations, suggesting that measurements made over a 1 min time course would reflect initial rates of the hexose uptake. To determine K<sub>m</sub> and V<sub>max</sub> of the endogenous glucose transporter(s) in Sf21 cells, the uptake of 2dGlc was measured over a range of substrate concentrations (50 μM~10 mM). 2dGlc uptake by the Sf21 cells appeared to involve both saturable and non-saturable (or very low affinity) components. A saturable transport system far 2dGlc was relatively high, the K<sub>m</sub> value for uptake being<0.45 mM. The V<sub>max</sub> value obtained for 2dGlc transport in the Sf21 cells was about 9.7-folds higher than that reported for Chinese hamster ovary cells, which contain a GLUT1 homologue. Thus, it appeared that the transport activity of the Sf21 cells was very high. In addition, the Sf21 glucose transporter was found to have very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter

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